Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling.
Author
Ast, JuliaNasteska, Daniela
Fine, Nicholas H F
Nieves, Daniel J
Koszegi, Zsombor
Lanoiselée, Yann
Cuozzo, Federica
Viloria, Katrina
Bacon, Andrea
Luu, Nguyet T
Newsome, Philip N
Calebiro, Davide
Owen, Dylan M
Broichhagen, Johannes
Hodson, David J
Publication date
2023-01-18
Metadata
Show full item recordAbstract
The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in glucose homeostasis and food intake. GLP1R agonists (GLP1RA) are widely used in the treatment of diabetes and obesity, yet visualizing the endogenous localization, organization and dynamics of a GPCR has so far remained out of reach. In the present study, we generate mice harboring an enzyme self-label genome-edited into the endogenous Glp1r locus. We also rationally design and test various fluorescent dyes, spanning cyan to far-red wavelengths, for labeling performance in tissue. By combining these technologies, we show that endogenous GLP1R can be specifically and sensitively detected in primary tissue using multiple colors. Longitudinal analysis of GLP1R dynamics reveals heterogeneous recruitment of neighboring cell subpopulations into signaling and trafficking, with differences observed between GLP1RA classes and dual agonists. At the nanoscopic level, GLP1Rs are found to possess higher organization, undergoing GLP1RA-dependent membrane diffusion. Together, these results show the utility of enzyme self-labels for visualization and interrogation of endogenous proteins, and provide insight into the biology of a class B GPCR in primary cells and tissue.Citation
Ast J, Nasteska D, Fine NHF, Nieves DJ, Koszegi Z, Lanoiselée Y, Cuozzo F, Viloria K, Bacon A, Luu NT, Newsome PN, Calebiro D, Owen DM, Broichhagen J, Hodson DJ. Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling. Nat Commun. 2023 Jan 18;14(1):301. doi: 10.1038/s41467-022-35716-1Type
ArticleAdditional Links
http://www.nature.com/ncomms/index.htmlPMID
36653347Journal
Nature CommunicationsPublisher
Nature Publishing Groupae974a485f413a2113503eed53cd6c53
10.1038/s41467-022-35716-1